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Direct Regulation of TLR5 Expression by Caveolin-1.

Lim JS, Nguyen KC, Han JM, Jang IS, Fabian C, Cho KA - Mol. Cells (2015)

Bottom Line: To assess the effect of caveolin-1 on TLR5, we analyzed TLR5 expression in senescent fibroblasts and aged tissues expressing high levels of caveolin-1.Moreover, overexpressed caveolin-1 in vitro enhanced TLR5 mRNA through the MAPK pathway and prolonged TLR5 protein half-life through direct interaction.These results suggest that caveolin-1 may play a crucial role in maintaining of TLR5 by regulating transcription systems and increasing protein half-life.

View Article: PubMed Central - PubMed

Affiliation: Department of Biochemistry, Chonnam National University Medical School, Gwangju 501-746, Korea.

ABSTRACT
Toll-like receptor 5 (TLR5) is a specific receptor for microbial flagellin and is one of the most well-known receptors in the TLR family. We reported previously that TLR5 signaling is well maintained during aging and that caveolin-1 may be involved in TLR5 signaling in aged macrophages through direct interactions. Therefore, it is important to clarify whether caveolin-1/TLR5 interactions affect TLR5 expression during aging. To assess the effect of caveolin-1 on TLR5, we analyzed TLR5 expression in senescent fibroblasts and aged tissues expressing high levels of caveolin-1. As expected, TLR5 mRNA and protein expression was well maintained in senescent fibroblasts and aged tissues, whereas TLR4 mRNA and protein were diminished in those cells and tissues. To determine the mechanism of caveolin-1-dependent TLR5 expression, we examined TLR5 expression in caveolin-1 deficient mice. Interestingly, TLR5 mRNA and protein levels were decreased dramatically in tissues from caveolin-1 knockout mice. Moreover, overexpressed caveolin-1 in vitro enhanced TLR5 mRNA through the MAPK pathway and prolonged TLR5 protein half-life through direct interaction. These results suggest that caveolin-1 may play a crucial role in maintaining of TLR5 by regulating transcription systems and increasing protein half-life.

No MeSH data available.


Related in: MedlinePlus

Caveolin-1 may be a crucial TLR5 mediator. Caveolin-1 was overexpressed in HeLa and NS-HDF cells using a lentivirus (LV) carrying RFP-caveolin-1 (RFP/Cav1). (A, B) Caveolin-1, TLR4, and TLR5 mRNA levels in caveolin-1 over-expressed HeLa and NS-HDF were analyzed by RT-PCR with specific primers, respectively. (C, D) Caveolin-1, TLR4, and TLR5 protein expression in caveolin-1 overexpressed HeLa and NS-HDF was detected by Western blotting with specific antibodies. The β-actin was used as the loading control for both RT-PCR and Western blotting. The relative density of protein expression and mRNA levels were normalized to β-actin and represented by quantitative graphs. Data are presented as mean ± SD from five independent experiments; *p < 0.05; ns, not significant.
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f4-molce-38-12-1111: Caveolin-1 may be a crucial TLR5 mediator. Caveolin-1 was overexpressed in HeLa and NS-HDF cells using a lentivirus (LV) carrying RFP-caveolin-1 (RFP/Cav1). (A, B) Caveolin-1, TLR4, and TLR5 mRNA levels in caveolin-1 over-expressed HeLa and NS-HDF were analyzed by RT-PCR with specific primers, respectively. (C, D) Caveolin-1, TLR4, and TLR5 protein expression in caveolin-1 overexpressed HeLa and NS-HDF was detected by Western blotting with specific antibodies. The β-actin was used as the loading control for both RT-PCR and Western blotting. The relative density of protein expression and mRNA levels were normalized to β-actin and represented by quantitative graphs. Data are presented as mean ± SD from five independent experiments; *p < 0.05; ns, not significant.

Mentions: To elucidate the function of caveolin-1 in the regulation of TLR5, caveolin-1 was overexpressed in NS-HDF and HeLa cells with low level of caveolin-1. Caveolin-1 over-expression increased TLR5 mRNA level in both cell types but not that of TLR4 (Fig. 4A). TLR5 protein expression was increased significantly in caveolin-1 overexpressed cells than that in control cells, while the TLR4 protein was not influenced by caveolin-1 overexpression (Fig. 4B). These results indicate that both TLR5 mRNA and protein were affected by exogenous caveolin-1 expression and that caveolin-1 may be a novel regulator of TLR5 to maintain the receptor during aging.


Direct Regulation of TLR5 Expression by Caveolin-1.

Lim JS, Nguyen KC, Han JM, Jang IS, Fabian C, Cho KA - Mol. Cells (2015)

Caveolin-1 may be a crucial TLR5 mediator. Caveolin-1 was overexpressed in HeLa and NS-HDF cells using a lentivirus (LV) carrying RFP-caveolin-1 (RFP/Cav1). (A, B) Caveolin-1, TLR4, and TLR5 mRNA levels in caveolin-1 over-expressed HeLa and NS-HDF were analyzed by RT-PCR with specific primers, respectively. (C, D) Caveolin-1, TLR4, and TLR5 protein expression in caveolin-1 overexpressed HeLa and NS-HDF was detected by Western blotting with specific antibodies. The β-actin was used as the loading control for both RT-PCR and Western blotting. The relative density of protein expression and mRNA levels were normalized to β-actin and represented by quantitative graphs. Data are presented as mean ± SD from five independent experiments; *p < 0.05; ns, not significant.
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4697003&req=5

f4-molce-38-12-1111: Caveolin-1 may be a crucial TLR5 mediator. Caveolin-1 was overexpressed in HeLa and NS-HDF cells using a lentivirus (LV) carrying RFP-caveolin-1 (RFP/Cav1). (A, B) Caveolin-1, TLR4, and TLR5 mRNA levels in caveolin-1 over-expressed HeLa and NS-HDF were analyzed by RT-PCR with specific primers, respectively. (C, D) Caveolin-1, TLR4, and TLR5 protein expression in caveolin-1 overexpressed HeLa and NS-HDF was detected by Western blotting with specific antibodies. The β-actin was used as the loading control for both RT-PCR and Western blotting. The relative density of protein expression and mRNA levels were normalized to β-actin and represented by quantitative graphs. Data are presented as mean ± SD from five independent experiments; *p < 0.05; ns, not significant.
Mentions: To elucidate the function of caveolin-1 in the regulation of TLR5, caveolin-1 was overexpressed in NS-HDF and HeLa cells with low level of caveolin-1. Caveolin-1 over-expression increased TLR5 mRNA level in both cell types but not that of TLR4 (Fig. 4A). TLR5 protein expression was increased significantly in caveolin-1 overexpressed cells than that in control cells, while the TLR4 protein was not influenced by caveolin-1 overexpression (Fig. 4B). These results indicate that both TLR5 mRNA and protein were affected by exogenous caveolin-1 expression and that caveolin-1 may be a novel regulator of TLR5 to maintain the receptor during aging.

Bottom Line: To assess the effect of caveolin-1 on TLR5, we analyzed TLR5 expression in senescent fibroblasts and aged tissues expressing high levels of caveolin-1.Moreover, overexpressed caveolin-1 in vitro enhanced TLR5 mRNA through the MAPK pathway and prolonged TLR5 protein half-life through direct interaction.These results suggest that caveolin-1 may play a crucial role in maintaining of TLR5 by regulating transcription systems and increasing protein half-life.

View Article: PubMed Central - PubMed

Affiliation: Department of Biochemistry, Chonnam National University Medical School, Gwangju 501-746, Korea.

ABSTRACT
Toll-like receptor 5 (TLR5) is a specific receptor for microbial flagellin and is one of the most well-known receptors in the TLR family. We reported previously that TLR5 signaling is well maintained during aging and that caveolin-1 may be involved in TLR5 signaling in aged macrophages through direct interactions. Therefore, it is important to clarify whether caveolin-1/TLR5 interactions affect TLR5 expression during aging. To assess the effect of caveolin-1 on TLR5, we analyzed TLR5 expression in senescent fibroblasts and aged tissues expressing high levels of caveolin-1. As expected, TLR5 mRNA and protein expression was well maintained in senescent fibroblasts and aged tissues, whereas TLR4 mRNA and protein were diminished in those cells and tissues. To determine the mechanism of caveolin-1-dependent TLR5 expression, we examined TLR5 expression in caveolin-1 deficient mice. Interestingly, TLR5 mRNA and protein levels were decreased dramatically in tissues from caveolin-1 knockout mice. Moreover, overexpressed caveolin-1 in vitro enhanced TLR5 mRNA through the MAPK pathway and prolonged TLR5 protein half-life through direct interaction. These results suggest that caveolin-1 may play a crucial role in maintaining of TLR5 by regulating transcription systems and increasing protein half-life.

No MeSH data available.


Related in: MedlinePlus